20S蛋白酶体β5亚单位对小鼠神经干细胞增殖和分化能力的影响  

Effects of 20S proteasome β5 subunit on proliferation and differentiation ability of mice neural stem cells

作　　者：周晓芳 王晓强 王婷玉 陆利[1] 赵云鹤[1] Zhou Xiaofang;Wang Xiaoqiang;Wang Tingyu;Lu Li;Zhao Yunhe(Department of Human Anatomy,Shanxi Medical University,Taiyuan 030001,China;The Second Clinical Hospital,Shanxi Medical University,Taiyuan 030001,China)

机构地区：[1]山西医科大学人体解剖学教研室,太原030001 [2]山西医科大学第二临床医学院,太原030001

出　　处：《神经解剖学杂志》2020年第3期253-260,共8页Chinese Journal of Neuroanatomy

基　　金：山西省青年科技研究基金(201801D221407);山西医科大学博士启动基金(03201604);山西医科大学大学生创新项目(20160232)。

摘　　要：目的:探讨20S蛋白酶体β5亚单位(PSMB5)对小鼠神经干细胞(NSCs)增殖和分化能力的影响.方法:体外分离培养新生BALB/c小鼠(P0)的NSCs,应用免疫荧光染色观察PSMB5在NSCs中的表达与分布;利用real time RT-PCR方法检测PSMB5 mRNA的表达;荧光分光光度法测定PSMB5相关的糜蛋白酶活性的变化;应用PSMB5-siRNA抑制P0 NSCs中PSMB5基因的表达;使用含有PSMB5基因的重组慢病毒((lenti-PSMB5))感染成年小鼠(P90)NSCs.利用CCK-8试验检测PSMB5对NSCs增殖能力的影响,利用BrdU和Tuj1染色观察PSMB5基因敲低和过表达对NSCs增殖及分化能力的影响.结果:PSMB5主要分布于NSCs的胞浆,P0的NSCs分化后,PSMB5 mRNA的表达下调.成年和老年小鼠NSCs中糜蛋白酶体活性较新生小鼠显著降低.PSMB5敲减后NSCs增殖能力和向神经元分化能力减弱,PSMB5过表达后NSCs增殖能力和向神经元分化能力增强.结论:PSMB5可能参与小鼠NSCs增殖及分化的调控.Objective: To investigate the effects of 20S subunit of proteasome subunit beta type-5(PSMB5) on the proliferation and differentiation of mice neural stem cells(NSCs).Methods: NSCs were isolated and cultured from BALB/c mice.The expression and localization of PSMB5 were observed by immunofluorescence staining.The expression of PSMB5 mRNA was detected by real time RT-PCR.The changes of PSMB5-related chymotrypsin activity was detected by fluorescence spectrophotometry.The expression of PSMB5 was inhibited by PSMB5-siRNA in P0 NSCs.And the expression of PSMB5 was upregulated by PSMB5 recombinant lentivirus(lenti-PSMB5) infection in adult mice(P90) NSCs.The effects of PSMB5 on NSCs proliferation were detectd by CCK-8 kit.The effects of PSMB5 on NSCs differentiation were detected by BrdU and Tuj1 staining.Results: PSMB5 was mainly distributed in the cytoplasm of NSCs.After differentiation of NSCs with P0,the expression of PSMB5 mRNA was down-regulated.The activity of chymotrypsin in NSCs of adult and aged mice was significantly lower than that of newborn mice.The proliferation and neuronal differentiation of NSCs were inhibited after PSMB5 knockdown,while the proliferation and neuronal differentiation of NSCs were enhanced after PSMB5 overexpression.Conclusion: PSMB5 may be involved in the regulation of proliferation and differentiation of mice NSCs.

关 键 词：神经干细胞 20S蛋白酶体亚β5单位 增殖 分化 小鼠 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]